Biden, speaking to about 60 US business leaders in the Chinese capital, expressed alarm over the country's move to expand its military influence in the skies over the East China Sea, and said he discussed the issue during more than four hours of talks with Chinese President Xi Jinping.
"China's recent and sudden announcement of the establishment of a new air defence identification zone has, to state the obvious, caused significant apprehension in the region," Biden said.
"And I was very direct about our firm position and our expectations in my conversations with President Xi."
China declared last month that it had established an air defence identification zone (ADIZ), over an area off its southeast coast that includes a group of islands disputed with Japan.
It says all aircraft within it must obey its instructions or risk unspecified "defensive emergency measures".
The move provoked anger in the region and prompted the US to defy Beijing by flying two B-52 bombers into the newly-declared zone.
Biden's remarks came on the final day of his two-day visit to Beijing.
The issue has dominated the agenda of his trip, which took him to Tokyo earlier this week and was due to carry on in Seoul later Thursday.
Neither Biden nor Xi made any direct mention of the ADIZ in their comments to reporters after Wednesday's meetings.
But a senior White House official reiterated that while the US did not recognise China's move, it was key that Beijing did not take any actions connected to the zone that would escalate the situation.
"We also made clear that not just the United States, but other countries as well are looking at them to take steps to lower tensions, and that includes avoiding enforcement actions that really could lead to a crisis," said the official, who requested anonymity in order discuss the details of the talks.
Beijing's foreign ministry said in a statement Thursday that at the meetings the Chinese side "stressed that China's move is in accordance with international law and conventions and that the American side should take an objective and fair attitude".
Beijing sees Tokyo as the aggressor in the dispute, which the state-run Global Times newspaper argued in an editorial has been "stirred up by Japan to win pity".
"Japan deliberately created a crisis and tried to get the US involved, hoping the US and China will evolve into confrontation," the newspaper wrote, adding that "it would be a nightmare" if the US and China were to "actively confront each other".
Despite the heightened nationalistic feelings and rhetoric surrounding the issue -- which hinges on a decades-old dispute over the island group, which Beijing calls Diaoyu and Tokyo, which controls them, calls Senkaku -- experts say that any outbreak of violence remains unlikely, as the major trade relationship between the two Asian powers is a strong incentive for both sides to avoid conflict.
In his remarks Thursday, Biden described a "complex" US-China relationship and acknowledged that "we have our differences and they are real".
"But there's nothing inevitable about a conflict with China -- nothing inevitable about a conflict with China," he added. "Wholesome competition and strong competition is fundamentally different than conflict."
Quoting Irish poet William Butler Yeats' work Easter 1916 which talks of a "terrible beauty" being born during Ireland's rebellion against British rule a century ago, he said it better describes the Pacific basin in 2013.
"'All's changed, changed utterly'," he said. "We're at a moment, a moment -- a window, as they say -- of opportunity, and how long it will remain open remains to be seen."
There was a chance to establish "a set of rules of the road that provide for mutual benefit and growth (for) both our countries and the region... for progress in the 21st century," Biden said.